水稻非生物胁迫响应基因OsMIP1的表达与进化分析

doi:10.11983/CBB16081

植物学报 ›› 2017, Vol. 52 ›› Issue (1): 43-53.DOI: 10.11983/CBB16081

所属专题：水稻生物学专辑 (2017年52卷1期)

水稻非生物胁迫响应基因OsMIP1的表达与进化分析

王玲^1,², 郭长奎^1,^2,³, 任丁^1,^2,^*(), 马红^1,^2,^*()

¹复旦大学生命科学学院, 上海 200438
²复旦大学植物科学研究所, 上海 200438
³浙江农林大学农业与食品科学学院, 杭州 311300

收稿日期:2016-04-16 接受日期:2016-05-26 出版日期:2017-01-01 发布日期:2017-01-23
通讯作者: 任丁,马红
作者简介:
# 共同第一作者
基金资助:
转基因生物新品种培育(No.2014ZX08009-014B);转基因生物新品种培育(No.2014ZX08009-014B)

Molecular Evolution and Expression Analysis of the OsMIP1 Response to Abiotic Stress

Wang Ling^1,², Guo Changkui^1,^2,³, Ren Ding^1,^2,^*(), Ma Hong^1,^2,^*()

¹School of Life Sciences, Fudan University, Shanghai 200438, China
²Institute of Plant Biology, Fudan University, Shanghai 200438, China
³School of Agriculture and Food Science, Zhejiang Agriculture and Forestry University, Hangzhou 311300, China

Received:2016-04-16 Accepted:2016-05-26 Online:2017-01-01 Published:2017-01-23
Contact: Ren Ding,Ma Hong
About author:
# Co-first authors

摘要/Abstract

摘要： MID1编码R-R型的MYB转录因子, 对不同的非生物胁迫均有响应, 特别是在水稻(Oryza sativa)生殖期会受到干旱胁迫的诱导, 进而在一定程度上可以保持花粉的育性并稳定水稻产量。为进一步研究水稻MID1对非生物胁迫的响应网络, 利用酵母双杂交系统筛选出与其互作的蛋白因子OsMIP1, 并利用双分子荧光互补系统在本氏烟草(Nicotiana benthamiana)细胞中得到验证。结果表明, OsMIP1编码1个预测含有ENTH/ANTH/VHS结构域的跨膜转运蛋白。OsMIP1在根、茎、叶、小穗和胚乳中均有表达。干旱胁迫下, OsMIP1在叶片和生殖器官中表达, 特别是在减数分裂后的小花中表达显著上调。这些结果暗示, OsMIP1在花器官抵抗干旱胁迫中起一定的作用。在水稻营养生长阶段, OsMIP1表达还受到包括NaCl和甘露醇在内的其它非生物胁迫的影响, 暗示其可能在其它非生物胁迫调节中也具有一定的作用。植物中关于编码ENTH/ ANTH/VHS结构域蛋白的研究很少。通过对MIP1亚家族进化关系进行分析, 结果表明, 在被子植物中, MIP1可分为6大类, 这6大类分别来自被子植物祖先中原本就存在的6个拷贝, 在被子植物的进化过程中又经历了多次基因重复和拷贝丢失等事件。MIP1家族成员广泛分布于被子植物中并可能具有抗胁迫等功能。

Abstract: The gene MID1 (MYB IMPORTANT FOR DROUGHT RESPONSE1) encodes a putative R-R type MYB transcription factor; is induced by abiotic stresses, especially drought in reproductive stage; and can improve pollen fertility and rice production. To understand the role of MID1 in abiotic stress responses, we used the yeast two-hybrid system to find an interacting protein, OsMIP1 (Oryza sativa MID1 interaction protein 1). The interaction was further confirmed by BiFC (bimolecular fluorescence complementation) analysis in tobacco leaf cells. OsMIP1 encodes a putative transmembrane protein with an ENTH/ANTH/VHS domain. It is expressed in the root, stem, leaf, panicle and endosperm. Under drought stress, its expression is upregulated in leaf and reproductive organs, especially in post-meiotic flowers. OsMIP1 may play a role in response to drought stress during reproductive development. OsMIP1 expression during vegetative development can be induced by other abiotic stress, including NaCl and mannitol, which suggests that OsMIP1 can respond to other abiotic stresses. There is little analysis of the evolution of genes encoding proteins with the ENTH/ ANTH/VHS domain, so we analyzed the molecular evolution of MIP1 homologs in flowering plants. The evolution analysis of the MIP1 family in angiosperms showed that MIP1 homologs can be divided into 6 types, which originated from at least 6 copies of MIP1 homologous genes in the ancestor of extent angiosperms. After gene-duplication and -loss events, MIP1 family members widely distributed in the angiosperms and might have various functions, possibly in stress responses.

王玲, 郭长奎, 任丁, 马红. 水稻非生物胁迫响应基因OsMIP1的表达与进化分析. 植物学报, 2017, 52(1): 43-53.

Wang Ling, Guo Changkui, Ren Ding, Ma Hong. Molecular Evolution and Expression Analysis of the OsMIP1 Response to Abiotic Stress. Chinese Bulletin of Botany, 2017, 52(1): 43-53.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB16081

https://www.chinbullbotany.com/CN/Y2017/V52/I1/43

图/表 10

图1 OsMIP1蛋白质的保守结构域预测

Figure 1 Analysis of conserved domains of OsMIP1 protein

图2 OsMIP1与MID1相互作用 (A) 在酵母中OsMIP1与MID1相互作用; (B) 在烟草叶片中, 利用双分子荧光互补验证OsMIP1与MID1相互作用(YFP: 黄色荧光蛋白; Merge: YFP和亮场2个图像的合并图像)。

Figure 2 OsMIP1 interacts with MID1 (A) OsMIP1 interacts with MID1 in yeast; (B) BiFC (bimole- cular fluorescence complementation) analysis of OsMIP1- MID1 interaction in tobacco leaf (YFP: Yellow fluorescent protein; Merge: Merged image)

图3 OsMIP1的亚细胞定位YFP: 黄色荧光蛋白; Merge: YFP和亮场2个图像的合并图像

Figure 3 Subcellular localization of OsMIP1YFP: Yellow fluorescent protein; Merge: Merged image

图4 OsMIP1在不同组织中的表达模式R: 根; S: 茎; L: 叶; MF: 减数分裂时期的小花; PMF: 减数分裂后的小花; DS: 灌浆5天的种子

Figure 4 Expression patterns of OsMIP1 in different tissuesR: Root; S: Stem; L: Leaf; MF: Meiotic floret; PMF: Post- meiotic floret; FS: 5-day-filling seeds

图5 OsMIP1在不同非生物胁迫下的表达模式(A) OsMIP1在干旱胁迫下不同组织中的表达模式; (B) OsMIP1在200 mmol·L-1甘露醇、20% PEG和200 mmol·L-1 NaCl胁迫下的表达模式。SL: 苗期叶片; MF: 减数分裂时期的小花; PL: 内稃和外稃; P: 雌蕊; S: 雄蕊; DS: 灌浆5天的种子

Figure 5 Expression patterns of OsMIP1 in different abiotic stress(A) Expression patterns of OsMIP1 in different tissues under drought; (B) Expression patterns of OsMIP1 in 200 mmol·L-1 mannitol, 20% PEG and 200 mmol·L-1 NaCl stress. SL: Seedling leaf; MF: Meiotic floret; PL: Palea and lemma; P: Pistil; S: Stamen; FS: 5-day filling seeds

表1 基因组数据中选择的物种及信息

Table 1 Species from genome database and their information

Class	Order	Species	Numbers
Eudicots	Cucurbitales	Cucumis sativus	5
	Fabales	Glycine max	16
	Malvales	Theobroma cacao	6
	Myrtales	Eucalyptus robusta	8
	Brassicales	Carica papaya	3
	Ranunculales	Aquilegia viridiflora	7
	Vitales	Vitis vinifera	7
	Brassicales	Arabidopsis thaliana	9
	Malpighiales	Populus trichocarpa	12
	Lamiales	Mimulus luteus	9
	Solanales	Solanum tuberosum	1
Monocots	Poales	Zea mays	14
	Poales	Brachypodium distachyon	10
	Poales	Oryza sativa	10
	Poales	Panicum virgatum	16
	Poales	Setaria italica	9
	Poales	Sorghum bicolor	10
ANA grade	Amborellales	Amborella trichopoda	7

表2 转录组数据中选择的物种及信息

Table 2 Species from transcriptome database and their in- formation

Class	Order	Species	Numbers
Eudicots	Apiales	Hedera nepalensis	9
	Aquifoliales	Ilex chinensis	8
	Asterales	Lactuca sativa	7
	Celastrales	Euonymus carnosus	9
	Cornales	Swida wilsoniana	9
	Dipsacales	Lonicera japonica	7
	Ericales	Camellia japonica	11
	Fagales	Cyclobalanopsis glauca	6
	Cornales	Aucuba japonica	8
	Gentianales	Vinca major	8
	Proteales	Platanus occidentalis	5
	Oxalidales	Oxalis corniculata	7
	Proteales	Meliosma arviflora	12
	Sapindales	Buxus sinica	6
Monocots	Alismatales	Acorus calamus	6
	Asparagales	Asparagus officinalis	4
	Asparagales	Phalaenopsis amabilis	6
	Liliales	Dioscorea opposita	8
	Liliales	Yucca smalliana	6
	Pandanales	Pandanus tectorius	5
	Zingiberales	Canna indica	8

图6 被子植物MIP1基因家族邻接/最大似然(NJ/ML)进化树被子植物中, MIP1基因可分为6大类(亚家族); 红色: 单子叶植物; 紫色: 真双子叶植物; 绿色: 无油樟。分支上的数字分别表示邻接(neighboring joining, NJ)和最大似然(maximum likelihood, ML)树自展分析的支持率。星号代表自展支持率为100%; 减号表示支持率低于50%。枝长无意义。

Figure 6 The NJ/ML (neighbor joining/maximum likelihood) tree of MIP1 genes family in angiospermsThe MIP1 genes can be divided into six types (subfamilies) in angiosperm. Red: Monocotyledons; Purple: Eudicotyledons; Green: Amborella trichopoda. The numbers on each branch are bootstrap support values of NJ/ML (neighbor joining/maximum likelihood) trees. Asterisks stand for bootstrap of 100% and minus stand for bootstrap below 50%. The length of each branch is meaningless.

图7 简化的I类MIP1基因进化树红色: 双子叶植物; 紫色: 单子叶植物

Figure 7 Simplified gene tree of type I MIP1 homologsRed: Dicotyledons; Purple: Monocotyledons

图8 简化的V类MIP1基因进化树红色: 菖蒲; 紫色: 双子叶植物; 绿色: 无油樟

Figure 8 Simplified gene tree of type V MIP1 homologsRed: Acorus calamus; Purple: Dicotyledons; Green: Amborella trichopoda

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水稻非生物胁迫响应基因OsMIP1的表达与进化分析

Molecular Evolution and Expression Analysis of the OsMIP1 Response to Abiotic Stress

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